Os10g0445400 - Revision history

Rice2012: /* References */

2015-06-13T08:48:37Z

‎References

Lmw2015: /* Structured Information */

2015-06-12T04:00:32Z

‎Structured Information

Chengtao at 15:39, 19 May 2014

2014-05-19T15:39:12Z

Sw at 15:37, 19 May 2014

2014-05-19T15:37:56Z

Sw at 15:19, 19 May 2014

2014-05-19T15:19:33Z

Sw at 15:03, 19 May 2014

2014-05-19T15:03:16Z

Chengtao: /* Expression */

2014-05-19T14:45:43Z

‎Expression

Chengtao: /* References */

2014-05-19T14:43:54Z

‎References

Lymzzu2009 at 14:12, 19 May 2014

2014-05-19T14:12:58Z

Jiaohzh: /* References */

2014-05-19T14:06:26Z

‎References

← Older revision		Revision as of 08:48, 13 June 2015
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	[[Category:Japonica Chromosome 10]]		[[Category:Japonica Chromosome 10]]
	[[Category:Chromosome 10]]		[[Category:Chromosome 10]]
		+	== Structured Information ==

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 </references>
 [[Category:Genes]]
 [[Category:Japonica mRNA]]

← Older revision		Revision as of 15:39, 19 May 2014
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	A Y2H screen was performed to identify 6 proteins that interact with OsHCI1. To confirm these positive interactions with OsHCI1, full-length coding sequences of the top six genes, which exhibited strong α-galactosidase activity, were cloned into GAL4 activation domain, respectively. Full-length OsHCI1 and each interacting protein were co-transformed into the Y2H Gold strain and grown on QDO/X/A medium(Fig.2).		A Y2H screen was performed to identify 6 proteins that interact with OsHCI1. To confirm these positive interactions with OsHCI1, full-length coding sequences of the top six genes, which exhibited strong α-galactosidase activity, were cloned into GAL4 activation domain, respectively. Full-length OsHCI1 and each interacting protein were co-transformed into the Y2H Gold strain and grown on QDO/X/A medium(Fig.2).

		+	[[File:图片2.jpg]]

		+	Fig2.Identification of OsHCI1 interaction with six proteins. The full-length OsHCI1 was cloned into pGBKT7, and full-length OsPSA7 (Os01g59600, 20S proteasome subunitα7), OsPGLU1 (Os03g53800, periplasmic beta-glucosidase), OsbHLH065, (Os04g41570, basic/Helix-Loop-Helix transcription factor), OsGRP1 (Os05g02770, glycine-rich cell wall structural protein), OsPOX1 (Os07g48020, peroxidase), and Os14-3-3 (Os11g34450, 14-3-3 protein) were cloned into pGADT7. The combination of indicated constructs was co-transformed into the Y2H Gold yeast strain. Yeast cells were dropped onto DDO and QDO/X/A medium, and grown for 5 d separately to test protein-protein interactions. Supplemented BD-murine p53 with AD-SV 40 large T- antigen were used as positive controls (PC). BD-lamin and AD-SV 40 large T-antigen combinations were used as negative controls (NC).

← Older revision		Revision as of 15:37, 19 May 2014
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	A Y2H screen was performed to identify 6 proteins that interact with OsHCI1. To confirm these positive interactions with OsHCI1, full-length coding sequences of the top six genes, which exhibited strong α-galactosidase activity, were cloned into GAL4 activation domain, respectively. Full-length OsHCI1 and each interacting protein were co-transformed into the Y2H Gold strain and grown on QDO/X/A medium(Fig.2).		A Y2H screen was performed to identify 6 proteins that interact with OsHCI1. To confirm these positive interactions with OsHCI1, full-length coding sequences of the top six genes, which exhibited strong α-galactosidase activity, were cloned into GAL4 activation domain, respectively. Full-length OsHCI1 and each interacting protein were co-transformed into the Y2H Gold strain and grown on QDO/X/A medium(Fig.2).

−	~~[[File:图片2.jpg]]~~

−	Fig2.Identification of OsHCI1 interaction with six proteins. The full-length OsHCI1 was cloned into pGBKT7, and full-length OsPSA7 (Os01g59600, 20S proteasome subunitα7), OsPGLU1 (Os03g53800, periplasmic beta-glucosidase), OsbHLH065, (Os04g41570, basic/Helix-Loop-Helix transcription factor), OsGRP1 (Os05g02770, glycine-rich cell wall structural protein), OsPOX1 (Os07g48020, peroxidase), and Os14-3-3 (Os11g34450, 14-3-3 protein) were cloned into
−	pGADT7.The combination of indicated constructs was co-transformed into the Y2H Gold yeast strain. Yeast cells were dropped onto DDO and QDO/X/A medium, and grown for 5 d separately to test protein-protein interactions. Supplemented BD-murine p53 with AD-SV 40 large T- antigen were used as positive controls (PC). BD-lamin and AD-SV 40 large T-antigen combinations were used as negative controls (NC).

← Older revision		Revision as of 15:19, 19 May 2014
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	[[File:图片2.jpg]]		[[File:图片2.jpg]]

−	Fig2.Identification of OsHCI1 interaction with six proteins.	+	Fig2.Identification of OsHCI1 interaction with six proteins. The full-length OsHCI1 was cloned into pGBKT7, and full-length OsPSA7 (Os01g59600, 20S proteasome subunitα7), OsPGLU1 (Os03g53800, periplasmic beta-glucosidase), OsbHLH065, (Os04g41570, basic/Helix-Loop-Helix transcription factor), OsGRP1 (Os05g02770, glycine-rich cell wall structural protein), OsPOX1 (Os07g48020, peroxidase), and Os14-3-3 (Os11g34450, 14-3-3 protein) were cloned into
		+	pGADT7.The combination of indicated constructs was co-transformed into the Y2H Gold yeast strain. Yeast cells were dropped onto DDO and QDO/X/A medium, and grown for 5 d separately to test protein-protein interactions. Supplemented BD-murine p53 with AD-SV 40 large T- antigen were used as positive controls (PC). BD-lamin and AD-SV 40 large T-antigen combinations were used as negative controls (NC).
		+

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 [[File:图片1.jpg]]
-Fig.1. Expression levels of OsHCl1 in rice plants subjected to four abiotic stresses and four hormonal treatments.
+Fig.1. Expression levels of OsHCl1 in rice plants subjected to four abiotic stresses and four hormonal treatments.  Rice seedlings were subjected to abiotic stresses and plant hormone treatments. (A), heat (45 °C), cold (4 °C), NaCl (250mM), and dehydration (dehydration on two pieces of tissues paper); OsHsp90-1, LIP19, OsSalT, and OsbZIP23 were used as reliable stress-inducible genes for each abiotic stress treatment, respectively. (B) 0.1mM abscisic acid (ABA), 0.1mM jasmonic acid (JA), 1mM salicylic acid (SA), 50 μl l–1 ethylene (Ethyl); OsSalT, OsPBZ1, OsPR1b, and OsERF3, were used as reliable genes for each hormonal treatment, respectively. C and T indicate untreated control and stress-treated samples, respectively. The experiments were performed with three biological replicates.
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 [[File:图片3.jpg]]
-Fig3. Expression patterns of the response of interacting protein genes with OsHCI1 under heat treatment.
+Fig3. Expression patterns of the response of interacting protein genes with OsHCI1 under heat treatment. Two-week-old rice seedlings were exposed to basal (A) or acquired heat stress (B) and then placed to normal temperature for 2h. Each leaf sample was harvested at different time points.
 ===Localization ===
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-[[File:图片4.jpg]]                                                                                 [[File:图片5.jpg]]
+[[File:图片4.jpg]]
-Fig. 4. Subcellular localization of six interacting proteins.                       Fig.5. BiFC assay for six substrate proteins confirms the interaction with OsHCI1 in living cells.
+Fig. 4. Subcellular localization of six interacting proteins. The full-length OsPSA7 (A), OsPGLU1 (B), OsbHLH065 (C), OsGRP1 (D), OsPOX1 (E), Os14-3-3 (F), and empty EYFP (G) were tagged with DsRed2 and transiently expressed with p19 in Nicotiana leaves. Images were captured and merged by z-series optical sections after 5 days of agro-infiltration.
 ===Wild type vs. Mutant ===
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 [[File:图片6.jpg]]
-Fig. 6. Thermotolerance phenotype of 35S:OsHCI1-EYFP
+Fig. 6. Thermotolerance phenotype of 35S:OsHCI1-EYFP.  Seven-day-old seedlings of 35S:EYFP (empty vector, EV) and 35S:OsHCI1-EYFP T3 transgenic plants (three independent lines) were grown on agar in the light for 7d and heated to 38 °C for 90min, cooled 24 °C for 2h, then heated to 45 °C for 3h (acquired thermotolerance) or heated to 45 °C for 60min (basal thermotolerance). A, RT-PCR analysis of seven independent Col-0/35S:OsHCI1 T3 transgenic plants, control wild type (WT), and EV. (B) The phenotypes of EV and three independent OsHCI1-overexpressed plants were treated at various high temperatures. Images were captured 5 days after heat shock treatment. (C) Percentage of surviving plants relative to the control (EV) on the same plate was determined 5 days after heat shock. Bars indicate standard deviation from the mean over all experiments (n = 30). The experiments were performed with four biological replicates.

← Older revision		Revision as of 14:45, 19 May 2014
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−		+	This study also examined the expression patterns of the interacting partner genes with OsHCI1 under two different heat stresses via semi-quantitative RT-PCR with rice seedlings treated by basal or acquired heat shock treatments (Fig.3).These results suggest that heat shock results in high expression of the OsHCI1 transcript or protein, which can affect the transcript levels of its interacting genes.
−	These results suggest that heat shock results in high expression of the OsHCI1 transcript or protein, which can affect the transcript levels of its interacting genes.

	[[File:图片3.jpg]]		[[File:图片3.jpg]]

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 <ref name="pmid:15128028"> Larkindale J, Huang B. 2004. Thermotolerance and antioxidant systems in Agrostis stolonifera: involvement of salicylic acid, abscisic acid, calcium, hydrogen peroxide, and ethylene. Journal of Plant Physiology 161, 405–413. </ref>
 <ref name="pmid:15923322"> Larkindale J, Hall JD, Knight MR, Vierling E. 2005. Heat stress phenotypes of Arabidopsis mutants implicate multiple signaling pathways in the acquisition of thermotolerance. Plant Physiology 138, 882–897. </ref>
 </references>

← Older revision		Revision as of 14:12, 19 May 2014
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−	This study also examined the expression patterns of the interacting partner genes with OsHCI1 under two different heat stresses via semi-quantitative RT-PCR with rice seedlings treated by basal or acquired heat shock treatments (Fig.3). These results suggest that heat shock results in high expression of the OsHCI1 transcript or protein, which can affect the transcript levels of its interacting genes.	+	These results suggest that heat shock results in high expression of the OsHCI1 transcript or protein, which can affect the transcript levels of its interacting genes.

	[[File:图片3.jpg]]		[[File:图片3.jpg]]